Association Analysis of miR423 (rs6505162) and miR608 (rs4919510) Polymorphisms with Lung Cancer Risk

Abstract

Background and purpose: Lung cancer is one of the deadliest cancers in the world, early detection of lung cancer is one of the biggest challenges in treating this disease and can be very effective in saving patients. Considering the role of microRNAs in the development and progression of lung cancer, the effective factors in the expression and function of microRNAs, including the polymorphisms in the genes of microRNAs, can be used as biomarkers for the early diagnosis of lung cancer. This study aims to investigate the relationship between single nucleotide polymorphisms (rs4919510) of miR608 and (rs6505162) of miR-423 with the risk of lung cancer. Materials and methods: In this case-control study, genotypic analysis of rs6505162 miR-423 and rs4919510 miR-608 polymorphisms was performed on two groups including 110 lung cancer patients and 120 healthy individuals by PCR-RFLP method. After receiving the consent form from the patients, 5 ml of venous blood was collected from each of the study subjects, and the genomic DNA of each person was extracted using the salt precipitation method, the quality of the DNA was measured by absorption spectroscopy, and then all the samples were PCR And by using restriction enzyme digestion by Rsa I and PVUII enzymes, the genotype was determined for both polymorphisms. In the last step, the results of enzyme digestion were checked by electrophoresis on 2% agarose gel, to confirm the RFLP results from sequencing. Random samples were used. Statistical analysis of data was done with SPSS software. Results: In the investigation of mrs6505162 polymorphism in miR-423, the frequency distribution of CC, CA, and AA genotypes in lung cancer patients was 42.7, 34.5, and 22.7, respectively, and in the control group, 30.8, 54.2, 15, respectively which did not have a significant difference. The genotype CA compared to CC and also CA compared to CC+AA was related to the reduction of lung cancer (P=0.009, 95% CI: 0.256-0.829, OR=0.460; P=0.003, 95% CI: 0.262-0.760, OR=0.447) which can indicate the protective role of this genotype. In the investigation of rs4919510 polymorphism in miR608, the frequency of allele G was 16.4% in the patient group and 20.4% in the control group, and the frequency of allele C was 83.6% in the patient group and 79.6% in the control group. This allele frequency was not statistically significant with a probability ratio of 0.763 and a confidence interval of 1.227-0.474 (P=0.263). No significant difference was observed between the group of cancer patients and control subjects. Codominant, Dominant, Over dominant, and Recessive, statistically no significant difference was observed. Conclusion: According to our findings in miR-423, the CA genotype in rs6505162 polymorphism can have a protective role in lung cancer, but rs4919510 polymorphism in miR-608 was not significantly associated with lung cancer, further studies with more samples are suggested to confirm these findings.

Keywords

• microrna
• lung cancer
• polymorphism
• pcr-rflp
• restriction enzyme